Antibody-based diagnosis of small ruminant lentivirus infection in seminal fluid

Autor: Helena Crespo, B. San Román, Hugo Ramírez, Damián F. de Andrés, X. de Andrés, María Jesús Grilló, Idoia Glaria, A. Grandas, C. Goñi, Beatriz Amorena, Ramsés Reina, B. Hichou, L. García-Pastor, F. Quintín, S. Cianca, L.E. Vijil, M. M. Hernández
Přispěvatelé: Comisión Interministerial de Ciencia y Tecnología, CICYT (España), Nafarroako Gobernua, Universidad de Navarra
Rok vydání: 2009
Předmět:
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
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ISSN: 0093-691X
Popis: Ramírez, Hugo et al.
Antibody-based diagnosis of small ruminant lentiviruses (SRLVs) has been efficiently achieved using serum and milk, but not semen, for which polymerase chain reaction (PCR) has been proposed as a confirmatory technique. This work, involving 296 ovine (Ovis aries) and caprine (Capra hircus) semen donors, investigates whether seminal fluid (SF) can be reliably used in antibody-based SRLV diagnosis. First, a gold standard was established to assess the infection status and determine the sensitivity and specificity of three commercial enzyme-linked immunosorbent assays (ELISAs) in serum testing using Western blot and PCR as confirmatory tests. For SF testing, both gold standard and serum testing results were used as reference. The performance of SF testing was affected not only by the ELISA assay sensitivity (related to antigen spectrum) compared with that of the gold standard (as it occurred in serum testing) but also by SF sample quality and SF working dilution. Nonturbid SF samples, commonly collected in artificial insemination centers (AICs), were required. Compared with serum, SF testing had a decreased sensitivity in two of the ELISA assays (with original serum working dilutions ≤1/20 in serum testing) but reached a similar sensitivity (and specificity) in the assay designed to work at the highest serum dilution (1/500). A SF concentration of about 1/2 (250-fold that used in serum testing) was found optimal in this assay, yielding highly repeatable results that were in almost perfect agreement with those of serum testing (κ ± SE, 0.91 ± 0.81). Thus, SF ELISA can be reliably applied in antibody-based SRLV diagnosis. This information may be useful to control infection in AICs and animal and semen trade programs requiring health-certified quality of semen donors. © 2009 Elsevier Inc. All rights reserved.
This study was supported by grants from Spanish CICYT (AGL2006-13410-C06-01/GAN) and the Government of Navarra Department of Agriculture (2003-2005). H. Ramírez was supported by the PASPA program, UNAM, and the Public University of Navarra. I. Glaria was supported by the Government of Navarra, Department of Agriculture. We are thankful to P. Lana, C. Santamaría (ITG-Ganadero), the Government of Navarra, and to AICs and farmers, particularly from Navarra and Aragon, as well as CAPRIGAN Breeder Association for kindly providing samples involved this study.
Databáze: OpenAIRE
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